CN114828900A

CN114828900A - High efficiency myeloperoxidase activatable imaging agents

Info

Publication number: CN114828900A
Application number: CN202080086172.9A
Authority: CN
Inventors: 约翰·W·陈; 王翠华
Original assignee: General Hospital Corp
Current assignee: General Hospital Corp
Priority date: 2019-12-17
Filing date: 2020-12-16
Publication date: 2022-07-29
Also published as: IL293875A; US20230124429A1; EP4076542A1; CA3162412A1; KR20220114536A; JP2023507385A; WO2021127013A1; AU2020404988A1; BR112022009471A2

Abstract

Provided herein are compounds useful as imaging agents. The exemplary compounds provided herein are useful as myeloperoxidase imaging agents using magnetic resonance or nuclear imaging techniques. Also provided are methods for making the compounds provided herein and diagnostic methods of using the compounds.

Description

High efficiency myeloperoxidase activatable imaging agents

Federally sponsored research or development

The present invention was made with government support under grant numbers NS103998 and HL150305 awarded by the National Institutes of Health. The government has certain rights in this invention.

Cross reference to related applications

This application claims the benefit of U.S. provisional application serial No. 62/949,336, filed on 2019, 12, month 17, the disclosure of which is incorporated herein by reference in its entirety.

Technical Field

The present invention relates to compounds useful as imaging agents, and more particularly to compounds useful as myeloperoxidase imaging agents.

Background

Myeloperoxidase (MPO) is a proinflammatory enzyme expressed in neutrophils and type M1 microglia and macrophages, but not by anti-inflammatory type M2 microglia and macrophages (see, e.g., Swirski et al, J.Clin. invest., 2010,120: 2627-2634; Bradley et al, Blood (Blood), 1982,60: 618-622; and Scholz et al, Experimental hematology (exp. hematol.) (2004, 32: 270-276). Although MPO is an integral part of innate immunity, it also damages tissues when secretion is abnormal.

Disclosure of Invention

The present application provides, inter alia, a compound of formula I:

or a pharmaceutically acceptable salt thereof, wherein:

ring a is selected from the group consisting of: c _6-10 Aryl, 4-16 membered heterocycloalkyl, and 5-16 membered heteroaryl;

ring B is selected from the group consisting of: c _6-10 Aryl, 4-16 membered heterocycloalkyl, and 5-16 membered heteroaryl;

L ¹ selected from the group consisting of: NHC (O) L ² 、NHC(O)-L ² -C(O)NH、L ² -NHC(O)-L ² 、L ² -NHC(O)-L ² -NHC (O) and L ² -NHC(O)-L ² -NHC(O)-L ² -；

Each L ² Independently selected from the group consisting of: c _1-4 Alkylene radical, C _1-4 Alkyleneoxy and C _1-4 An alkenylene group;

R ¹ comprising a chelating group anda metal;

R ² and R ³ Each independently selected from the group consisting of: OR (OR) ^a 、C(O)R ^a And OC (O) R ^a ；

Each R ^a Independently selected from the group consisting of: h and C _1-4 An alkyl group;

m is 0,1, 2, 3 or 4; and is

n is 0,1, 2, 3 or 4.

In some embodiments, ring a is selected from the group consisting of: phenyl, bicyclic 8-16 membered heterocycloalkyl, tricyclic 8-16 membered heterocycloalkyl, bicyclic 8-16 membered heteroaryl and tricyclic 8-16 membered heteroaryl. In some embodiments, ring a is selected from the group consisting of:

and

wherein

Represents rings A and L ¹ A bond between them.

In some embodiments, m is 1 or 2. In some embodiments, m is 1.

In some embodiments, each R is ² Independently selected from the group consisting of: OH, OCH ₃ 、C(O)CH ₃ And OC (O) CH ₃ . In some embodiments, each R is ² Is OH.

In some embodiments, ring a is selected from the group consisting of:

and

wherein

Represents rings A and L ¹ A bond between them.

In some embodiments, ring a is selected from the group consisting of:

and

wherein

Represents rings A and L ¹ A bond between them.

In some embodiments, ring B is selected from the group consisting of: phenyl, bicyclic 8-16 membered heterocycloalkyl, tricyclic 8-16 membered heterocycloalkyl, bicyclic 8-16 membered heteroaryl and tricyclic 8-16 membered heteroaryl. In some embodiments, ring B is selected from the group consisting of:

and

wherein

Represents rings B and L ¹ A bond between them.

In some embodiments, n is 1 or 2. In some embodiments, n is 1.

In some embodiments, each R is ³ Independently selected from the group consisting of: OH, OCH ₃ 、C(O)CH ₃ And OC (O) CH ₃ . In some embodiments, each R is ³ Is OH.

In some embodiments, ring B is selected from the group consisting of:

and

wherein

Represents rings B and L ¹ A bond between them.

In some embodiments, ring B is selected from the group consisting of:

and

wherein

Represents rings B and L ¹ A bond between them.

In some embodiments, ring a and ring B are the same. In some embodiments, ring a and ring B are different.

In some embodiments, ring a and ring B are eachIs composed of

In some embodiments, L ¹ Is L ² -NHC(O)-L ² -NHC(O)-L ² -, and each L ² Is independently selected C _1-4 An alkylene group.

In some embodiments, L ¹ Comprises the following steps:

wherein:

represents L ¹ A bond to ring a;

represents L ¹ A bond to ring B; and is provided with

Represents L ¹ And R ¹ A bond between them.

In some embodiments, R ¹ The metal of (a) is selected from the group consisting of: gd (Gd) ³⁺ 、Mn ²⁺ 、 ⁶⁸ Ga、 ⁶⁴ Cu and ¹¹¹ in. In some embodiments, R ¹ Said metal of (A) is Gd ³⁺ . In some embodiments, R ¹ The chelating group of (a) is selected from the group consisting of:

and

whereinM is said metal and

represents R ¹ And L ¹ A bond between them.

In some embodiments, R ¹ The method comprises the following steps:

wherein

Represents R ¹ And L ¹ A bond between them.

In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is a compound of formula II:

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is a compound of formula III:

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is a compound of formula IV:

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is a compound of formula Vb:

or a pharmaceutically acceptable salt thereof, wherein M is the metal.

In some embodiments, the compound of formula I is:

or a pharmaceutically acceptable salt thereof.

The present application further provides a pharmaceutical composition comprising a compound provided herein, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier.

The present application further provides a method of imaging a cell or tissue sample, the method comprising:

i) administering to a subject a compound provided herein or a pharmaceutically acceptable salt thereof;

ii) waiting for a time sufficient for the compound to accumulate at the cell or tissue sample; and

iii) imaging the cell or tissue sample using an imaging technique.

The present application further provides a method of imaging a liver of a subject, the method comprising:

ii) waiting for a time sufficient for the compound to accumulate at the liver; and

iii) imaging the cell or tissue sample using an imaging technique.

In some embodiments, the subject has been identified as having non-alcoholic steatohepatitis.

The present application further provides a method of diagnosing a disease or disorder associated with abnormal myeloperoxidase activity in a subject, the method comprising:

ii) waiting for a time sufficient for the compound to accumulate at a cellular or tissue site associated with the disease; and

iii) imaging the cell or tissue using imaging techniques.

In some embodiments, the method further comprises imaging the subject prior to step i).

The present application further provides a method of imaging myeloperoxidase activity in a cell, the method comprising:

i) contacting the cell with a compound provided herein or a pharmaceutically acceptable salt thereof; and

ii) imaging the cell with an imaging technique.

The present application further provides a method of detecting myeloperoxidase activity in a cell or tissue sample, the method comprising:

i) contacting the cell or tissue sample with a compound provided herein or a pharmaceutically acceptable salt thereof; and

ii) imaging the cell or tissue sample with an imaging technique.

The present application further provides a method of detecting myeloperoxidase activity in a subject, the method comprising:

i) administering to the subject a compound provided herein or a pharmaceutically acceptable salt thereof; and

ii) imaging the subject with an imaging technique.

The present application further provides a method of monitoring treatment of a disease or disorder associated with abnormal myeloperoxidase activity in a subject, the method comprising:

ii) imaging the subject with an imaging technique;

iii) administering to the subject a therapeutically effective amount of a therapeutic compound to treat the disease or disorder;

iv) imaging the cells or tissues of the subject with an imaging technique; and

v) comparing the image of step i) with the image of step iv).

In some embodiments, the method further comprises administering to the subject a compound provided herein, or a pharmaceutically acceptable salt thereof, after the administering of step iii) and before the imaging of step iv).

In some embodiments, the imaging technique is selected from the group consisting of magnetic resonance imaging and nuclear imaging.

In some embodiments, the disease or disorder associated with abnormal myeloperoxidase activity is selected from the group consisting of: non-alcoholic steatohepatitis, cancer, rheumatic diseases, infectious diseases, central nervous system diseases, cardiovascular disorders, autoimmune disorders, and inflammation associated with one or more of cancer, rheumatic diseases, infectious diseases, central nervous system diseases, cardiovascular disorders, and autoimmune disorders.

In some embodiments, the central nervous system disease is selected from the group consisting of: alzheimer's disease, stroke, epilepsy, Parkinson's disease, neurodegenerative diseases, and inflammation associated with one or more of Alzheimer's disease, stroke, epilepsy, Parkinson's disease, and neurodegenerative diseases.

In some embodiments, the cardiovascular disorder is selected from the group consisting of: atherosclerosis, myocardial infarction, atrial fibrillation, vasculitis, and inflammation associated with one or more of atherosclerosis, myocardial infarction, atrial fibrillation, and vasculitis.

In some embodiments, the autoimmune disorder is selected from the group consisting of: multiple sclerosis, meningitis, encephalitis, and inflammation associated with one or more of multiple sclerosis, meningitis, and encephalitis.

In some embodiments, the cancer is selected from the group consisting of: bladder cancer, breast cancer, carcinoma, cervical cancer, colorectal cancer, endometrial cancer, glioma, head and neck cancer, liver cancer, lung cancer, melanoma, ovarian cancer, pancreatic cancer, prostate cancer, renal cancer, skin cancer, stomach cancer, testicular cancer, leukemia, and thyroid cancer. In some embodiments, the cancer is a solid tumor.

In some embodiments, the rheumatic disease is selected from the group consisting of rheumatoid arthritis, osteoarthritis, and inflammatory arthritis.

In some embodiments, the inflammatory arthritis is selected from the group consisting of gout and calcium pyrophosphate deposition disease (CPPD).

In some embodiments, the infectious disease is selected from the group consisting of a fungal disease and a bacterial disease.

In some embodiments, the disease or disorder associated with abnormal myeloperoxidase activity is non-alcoholic steatohepatitis (NASH).

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials for use in the present invention are described herein; other suitable methods and materials known in the art may also be used. These materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

Drawings

FIG. 1 shows the chemical structures of a representative compound of formula I, mcMPO-Gd, and a previously reported compound, MPO-Gd.

FIG. 2A shows the T1 relaxivity of mcCPO-Gd measured at 0.33mM, 0.5mM, 0.75mM and 1mM in PBS at 0.47T and 40 ℃. Based on the 1/T1 value and the mcMPO-Gd concentration data (R) ² 0.996, n 3) the relaxation potency (relaxation) was calculated.

FIG. 2B shows the dynamic R1 ratio of mcMPO-Gd and MPO-Gd upon MPO activation. mcMPO-Gd showed faster activation compared to MPO-Gd when incubated with MPO, GOX and glucose for 180 minutes at 40 ℃.

Fig. 3 shows MTT assay results for RAW 264.7 cells. No significant toxicity was observed for mcMPO-Gd up to 5 mM.

FIGS. 4A-4D show mcMPO-Gd, DOTA-Gd, and MPO-Gd and Mpo in pre-contrast and wild type mice ^–/– Representative image at about 60 minutes of mcMPO-Gd in mice.

Fig. 4E shows the normalized contrast-to-noise ratio (CNR) of the MR imaging over a period of about 60 minutes.

Fig. 4F shows the activation ratio for MR imaging over a period of about 60 minutes, demonstrating that the activation ratio for mcMPO-Gd is about two times higher compared to MPO-Gd at about 60 minutes. Both compounds showed activation ratios much higher than DOTA-Gd and Mpo ^–/– Mouse mcMPO-Gd.

FIGS. 5A-5C show representative MR imaging of MPO-Gd and mcMPO-Gd. + -. AZM198 treatment in a serial stenosis model of atherosclerotic plaque instability.

Fig. 5D shows the contrast to noise ratio (CNR) of unstable plaques for MPO-Gd (0.3mmol/kg) and mcMPO-Gd (0.1mmol/kg) (n ═ 3 mice per probe), showing no significant difference even though the mcMPO-Gd was used at a dose 3-fold lower than MPO-Gd (p ═ 0.368, ns., not significant, determined by Mann-Whitney test).

Figure 5E shows Δ CNR of unstable and stable plaques (brachiocephalic stem) in control and AZM 198-treated TS mice. The signal in unstable plaques of AZM 198-treated mice was reduced by about 50% compared to the signal of no drug control (.: p ═ 0.039, determined by unpaired t-test). Δ CNR ═ CNR _{After the production of images} -CNR _{Before radiography} 。

Figure 6 shows CNR changes over 60 minutes in AZM198 treated mice and control mice. From 30 to 60 minutes, the signal from control mice increased, while the signal from AZM 198-treated mice decreased over this period.

FIG. 7 shows that intramolecular hydrogen bonding between the two amide bonds increases the rigidity of the mcMPO-Gd as shown by the molecular docking of the mcMPO-Gd bound to MPO.

FIG. 8 shows the binding affinity of mcMPO-Gd (left) and MPO-Gd (right) in the docking experiment described in example 13.

FIG. 9 shows a molecular docking representation of the binding of mcmPO-Gd (left) and MPO-Gd (right) to the MPO-CN complex using PyMol. Amino acid residues Q91, H95 and R239 (shown as lines) form together with heme (in stick form) the active site of MPO. All other amino acid residues (as indicated by the lines) are spaced from the mcMPO-Gd and MPO-Gd

Within. Hydrogen bonding and electrostatic interactions are shown in dashed lines.

Detailed Description

MPO is a factor in many diseases, including atherosclerosis (see, e.g., Teng et al, Redox. Rep.) 2017,22:51-73, vasculitis (see, e.g., Su et al, Radiology (Radiology) 2012,262:181-190), stroke (see, e.g., Forghani et al, J.Cereb. blood Flow Metab. 2015,35:485-493), Parkinson's disease (see, e.g., Choi et al, J.Neurosci.) -2005, 25:6594-6600), Alzheimer's disease (see, e.g., Ma et al, Biochem.). 2009,284:3158-3169, and Tka et al, Lekat. Chem. 198, J.Biochem.) -195-198, and Hakk.198, e.g., Ly., 2014. Shi. (Louis.) and Hakk et al, multiple sclerosis (see, e.g., Lekat. Redox. Rex. 2017,22:51-73), vasculitis, 190-190), and this enzyme is becoming a diagnostic and therapeutic target (see, e.g., Ruggeri et al, J.Med.chem.) -2015, 58:8513 8528; Juciate et al, Brain (Brain), 2015,138: 2687-2700; Malle et al, British J.Pharmacol. (Br.J.Pharmacol.) 2007,152: vala 854; and rashed et al, European Heart J.Heart J.) -2018, 39:3301-3310 previous reports describe an activatable MPO-specific gadolinium-based MRI agent bis-5-HT-Gd-DTPA (i.e., MPO-Gd; structures are shown in FIG. 1) (see, e.g., Rodriguez et al, American society of chemistry (J.Med.Som.63c.) -168: Gd-DTPA (i.e., MPO-Gd) (see, e.g., Rodriguez et al, J.Chem.J.Som.63c.) -168; Gd-27; and Brain (E) 112177) (see, E.E.M.11: 177) (see, E.E.M.M.3, Immunity et al, St., MDE., Atherosclerosis (see, e.g., Rashid et al, journal of european heart 2018,39: 3301-. MPO-Gd imaging also tracks stroke (see, e.g., Forghani et al, J. cerebrum flow and metabolism 2015,35: 485-), unstable atherosclerotic plaques (see, e.g., Rashid et al, J. European Heart 2018,39: 3301-.

However, the literature reports the concern of gadolinium deposition when using gadolinium-based contrast agents (GBCA), particularly linear GBCA, for Magnetic Resonance (MR) imaging based on the correlation between administration of GBCA and the development of nephrogenic systemic fibrosis in patients with renal failure (see, e.g., Grobner, t. "kidney disease dialysis transplant" (t. nephrol. dial.) -2006, 21: 1104-. Typical cases of gadolinium accumulation in renal systemic fibrosis and nervous tissue are associated with the use of linear GBCA such as gadolinium diamine (Omniscan), gadopentetate dimeglumine (Gd-DTPA or Magnevist) and OptiMARK. Recent studies have shown that gadolinium deposits following linear GBCA administration are also found in nerve tissue of patients and animals with normal renal function (see, e.g., McDonald et al, radiology, 2015,275: 772-.

Macrocyclic GBCAs bind gadolinium more tightly than linear GBCAs, thereby reducing the likelihood of dissociation of gadolinium ions from the chelating molecule. In line with this, administration of macrocyclic GBCA's, such as meglumine gadotetate (Dotarem) (see, e.g., Robert et al, radiology research 2015,50: 473-. Therefore, new imaging agents are needed to improve the safety and advantages of MR imaging while minimizing risks. Accordingly, the present application provides macrocyclic-based activatable MRI imaging agents for the detection of MPO activity.

Compound (I)

The present application provides compounds of formula I:

or a pharmaceutically acceptable salt thereof, wherein:

Each L ² Independently selected from the group consisting of: c _1-4 Alkylene radical, C _1-4 Alkylene radical, C _1-4 Alkyleneoxy and C _1-4 An alkenylene group;

R ¹ the method comprises the following steps: (i) a chelating group; or (ii) a chelating group and a metal;

m is 0,1, 2, 3 or 4; and is

n is 0,1, 2, 3 or 4.

In some embodiments, ring a is selected from the group consisting of: c _6-10 Aryl, bicyclic 8-16 membered heterocycloalkyl, tricyclic 8-16 membered heterocycloalkyl, bicyclic 8-16 membered heteroaryl and tricyclic 8-16 membered heteroaryl. In some embodiments, ring a is selected from the group consisting of: phenyl, bicyclic 8-16 membered heterocycloalkyl, tricyclic 8-16 membered heterocycloalkyl, bicyclic 8-16 membered heteroaryl and tricyclic 8-16 membered heteroaryl. In some embodiments, ring a is phenyl. In some embodiments, ring a is a bicyclic 8-16 membered heterocycloalkyl. In some embodiments, ring a is tricyclic 8-16 membered heterocycloalkyl. In some embodiments, ring a is bicyclic 8-16 membered heteroaryl. In some embodiments, ring a is a tricyclic 8-16 membered heteroaryl.

In some embodiments, ring a is selected from the group consisting of:

and

wherein

Represents rings A and L ¹ A bond between them.

In some embodiments, ring a is

In some embodiments, m is 0,1, 2, 3. In some embodiments, m is 1 or 2. In some embodiments, m is 1. In some embodiments, m is 2. In some embodiments, m is 3. In some embodiments, m is 4.

In some embodiments, ring a is selected from the group consisting of:

and

wherein

Represents rings A and L ¹ A bond between them.

In some embodiments, ring a is

In some embodiments, each R is ² Independently selected from the group consisting of: OR (OR) ^a 、C(O)R ^a And OC (O) R ^a And each R is ^a Independently selected from the group consisting of: h and CH ₃ . In some embodiments, each R is ² Independently selected from the group consisting of: OH, OCH ₃ 、C(O)CH ₃ And OC (O) CH ₃ . In some embodiments, each R is ² Is OH. In some embodiments, each R is ² Is OCH ₃ . In some embodiments, each R is ² Is C (O) CH ₃ . In some embodiments, each R is ² Is OC (O) CH ₃ 。

In some embodiments, ring a is selected from the group consisting of:

and

wherein

Represents rings A and L ¹ A bond between them.

In some embodiments, ring B is selected from the group consisting of: c _6-10 Aryl, bicyclic 8-16 membered heterocycloalkyl, tricyclic 8-16 membered heterocycloalkyl, bicyclic 8-16 membered heteroaryl and tricyclic 8-16 membered heteroaryl. In some embodiments, ring B is selected from the group consisting of: phenyl, bicyclic 8-16 membered heterocycloalkyl, tricyclic 8-16 membered heterocycloalkyl, bicyclic 8-16 membered heteroaryl and tricyclic 8-16 membered heteroaryl. In some embodiments, ring B is phenyl. In some embodiments, ring B is a bicyclic 8-16 membered heterocycloalkyl. In some embodiments, ring B is a tricyclic 8-16 membered heterocycloalkyl. In some embodiments, ring B is a bicyclic 8-16 membered heteroaryl. In some embodiments, ring B is a tricyclic 8-16 membered heteroaryl.

In some embodiments, ring B is selected from the group consisting of:

and

wherein

Represents rings B and L ¹ A bond between them.

In some embodiments, ring B is

In some embodiments, n is 0,1, 2, 3. In some embodiments, n is 1 or 2. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.

In some embodiments, ring B is selected from the group consisting of:

and

wherein

Represents rings B and L ¹ A bond between them.

In some embodiments, ring B is

In some embodiments, each R is ³ Independently selected from the group consisting of: OR (OR) ^a 、C(O)R ^a And OC (O) R ^a And each R is ^a Independently selected from the group consisting of: h and CH ₃ . In some embodiments, each R is ³ Independently selected from the group consisting of: OH, OCH ₃ 、C(O)CH ₃ And OC (O) CH ₃ . In some embodiments, each R is ³ Is OH. In some embodiments, each R is ³ Is OCH ₃ . In some embodiments, each R is ³ Is C (O) CH ₃ . In some embodiments, each R is ³ Is OC (O) CH ₃ 。

In some embodiments, ring B is selected from the group consisting of:

and

wherein

Represents rings B and L ¹ A bond between them.

In some embodiments, ring B is

In some embodiments, ring a and ring B are the same. In some embodiments, ring a and ring B are different. In some embodiments, ring a and ring B are each

In some embodiments, L ¹ Is NHC (O) L ² . In some embodiments, L ¹ Is NHC (O) -L ² -C (O) NH. In some embodiments, L ¹ Is L ² -NHC(O)-L ² . In some embodiments, L ¹ Is L ² -NHC(O)-L ² -nhc (o). In some embodiments, L ¹ Is L ² -NHC(O)-L ² -NHC(O)-L ² -。

In some embodiments, each L ² Independently selected from C _1-4 Alkylene and C _1-4 Alkenylene group; in some embodiments, each L ² Is independently selected C _1-4 An alkylene group. In some embodiments, each L ² Is independently selected C _1-4 An alkylene oxide group. In some embodiments, each L ² Is independently selected C _1-4 An alkenylene group.

In some embodiments, L ¹ Comprises the following steps:

wherein:

represents L ¹ A bond to ring a;

represents L ¹ A bond to ring B; and is

Represents L ¹ And R ¹ A bond between them.

In some embodiments, R ¹ Including a chelating group. In some embodiments, R ¹ Including chelating groups and metals. In some embodiments, R ¹ Including chelating groups and metals suitable for imaging using imaging techniques (e.g., magnetic resonance imaging, nuclear imaging, etc.). In some embodiments, R ¹ Including chelating groups and metals suitable for magnetic resonance imaging. In some embodiments, R ¹ Including chelating groups and metals suitable for nuclear imaging.

In some embodiments, R ¹ The metal of (a) is selected from the group consisting of: gd (Gd) ³⁺ 、Mn ²⁺ 、 ⁶⁸ Ga、 ⁶⁴ Cu and ¹¹¹ in. In some embodiments, R ¹ Said metal of (A) is Gd ³⁺ 。

In some embodiments, R ¹ The chelating group of (a) is selected from the group consisting of: 1,4, 7-Triazacyclononanetriacetic acid (NOTA), 1,4,7, 10-tetraazacyclododecane-1, 4,7, 10-tetraacetic acid (DOTA), 1,4, 7-TriazacyclononaneAlkane-1-glutaric acid-4, 7-diacetic acid (NODAGA) and ethylenediaminetetraacetic acid (EDTA).

In some embodiments, R ¹ Selected from the group consisting of:

and

wherein M is said metal and

represents R ¹ And L ¹ A bond between them.

In some embodiments, R ¹ Selected from the group consisting of:

and

wherein M is a metal selected from the group consisting of: gd (Gd) ³⁺ 、Mn ²⁺ 、 ⁶⁸ Ga、 ⁶⁴ Cu and ¹¹¹ in, and

represents R ¹ And L ¹ A bond between them.

In some embodiments, R ¹ Comprises the following steps:

wherein M is said metal and

represents R ¹ And L ¹ A bond between them.

In some embodiments, R ¹ The method comprises the following steps:

represents R ¹ And L ¹ A bond between them.

In some embodiments, R ¹ The method comprises the following steps:

wherein

Represents R ¹ And L ¹ A bond between them.

In some embodiments:

ring a is selected from the group consisting of: phenyl, bicyclic 8-16 membered heterocycloalkyl, tricyclic 8-16 membered heterocycloalkyl, bicyclic 8-16 membered heteroaryl and tricyclic 8-16 membered heteroaryl;

ring B is selected from the group consisting of: phenyl, bicyclic 8-16 membered heterocycloalkyl, tricyclic 8-16 membered heterocycloalkyl, bicyclic 8-16 membered heteroaryl and tricyclic 8-16 membered heteroaryl;

L ¹ is L ² -NHC(O)-L ² -NHC(O)-L ² -；

Each L ² Is independently selected C _1-4 An alkylene group;

R ¹ including chelating groups and metals;

m is 0,1 or 2; and is

n is 0,1 or 2.

In some embodiments:

ring a is selected from the group consisting of:

and

ring B is selected from the group consisting of:

and

L ¹ is L ² -NHC(O)-L ² -NHC(O)-L ² -；

Each L ² Is independently selected C _1-4 An alkylene group;

R ¹ including chelating groups and metals;

R ² and R ³ Each is OR ^a ；

m is 0,1 or 2; and is

n is 0,1 or 2.

In some embodiments:

ring a is selected from the group consisting of:

and

ring B is selected from the group consisting of:

and

L ¹ is L ² -NHC(O)-L ² -NHC(O)-L ² -；

Each L ² Is independently selected C _1-4 An alkylene group;

R ¹ including chelating groups and metals;

R ² and R ³ Each is OR ^a ；

m is 0,1 or 2; and is

n is 0,1 or 2.

In some embodiments:

ring a and ring B are each independently selected from the group consisting of:

and

L ¹ is L ² -NHC(O)-L ² -NHC(O)-L ² -；

Each L ² Is independently selected C _1-4 An alkylene group; and is

R ¹ Selected from the group consisting of:

and

wherein M is a metal selected from the group consisting of: gd (Gd) ³⁺ 、Mn ²⁺ 、 ⁶⁸ Ga、 ⁶⁴ Cu and ¹¹¹ in. In some embodiments:

ring A and ring B are each

L ¹ Is L ² -NHC(O)-L ² -NHC(O)-L ² -；

Each L ² Is independently selected C _1-4 An alkylene group; and is

R ¹ Is composed of

Wherein ^M Is a metal selected from the group consisting of: gd (Gd) ³⁺ 、Mn ² ⁺ 、 ⁶⁸ Ga、 ⁶⁴ Cu and ¹¹¹ In。

or a pharmaceutically acceptable salt thereof, wherein the variable R ¹ 、R ² 、R ³ M and n are defined according to the definitions provided herein for the compounds of formula I.

or a pharmaceutically acceptable salt thereof, wherein the variable R ¹ 、R ² And R ³ Are defined according to the definitions provided herein for compounds of formula I.

or a pharmaceutically acceptable salt thereof, wherein the variable R ¹ Are defined according to the definitions provided herein for compounds of formula I.

In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is a compound of formula V:

or a pharmaceutically acceptable salt thereof, wherein M is a metal and is defined according to the definitions provided herein for compounds of formula I.

In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is a compound of formula Va:

In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is a compound of formula Vc:

In some embodiments, the compound of formula I is:

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of formula I is:

or a pharmaceutically acceptable salt thereof.

Synthesis of

The synthesis of representative compounds of formula I, mcMPO-Gd, is shown in schemes 1A-1B, using the following general reaction conditions: (a) NaNO ₂ ，NaBr，0.75M HBr，-15℃；(b)MgSO ₄ ，H ₂ SO ₄ t-BuOH, rt, 45% (for two steps))；(c)Boc ₂ O，K ₂ CO ₃ ，THF/H ₂ O (1/4), rt; (d) edc.hcl, HOBt, DMF, rt, 82% (for both steps); (e) TFA/DCM (1/2), 63%; f) k ₂ CO ₃ ，CH ₃ CN, reflux, 78%; g) TFA/DCM, rt; h) GdCl ₃ pH 5.5, rt, then overnight at 50 ℃; i) EDC.HCl, HOBt, NEt ₃ ，DMSO，25％。

Scheme 1A.

Scheme 1B.

Compound 1 was synthesized in two steps according to published procedures (see, e.g., International patent application No. WO 2005/122682; and Moumne et al, J.org.chem., 2006,71: 3332-3334). Intermediate 4 was synthesized by three steps, starting with Boc protection of 5-hydroxytryptophan, followed by coupling with 5-hydroxyindoleacetic acid using 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC) to afford compound 3, which was Boc deprotected to afford intermediate 4. The synthesis of intermediate 8 starts with macrocyclic compound 5 and compound 1 under basic conditions to give compound 6. Compound 6 is hydrolyzed under acidic conditions and then reacted with GdCl ₃ Chelation (see, e.g., Kielar et al, J.Am.chem.Soc.). 2010,132:7836-7837) to provide intermediate 8. Intermediate 8 was then coupled with intermediate 4 using EDC hydrochloride and hydroxybenzotriazole (HOBt) to provide the final product mcMPO-Gd. The structure of mcMPO-Gd was confirmed by high resolution mass spectrometry.

Compared to linear chelates and other macrocyclic-based chelates, DOTA was chosen as a representative chelating group due to its thermodynamic and kinetic stability (see, e.g., Port et al, "Biometals, 2008,21: 469-490). As previously reported, two 5-hydroxyindole units were selected as MPO activatable moieties to enhance retention (see, e.g., Rodriguez et al, J. Am. chem. Soc. 2010,132: 168-177). As shown in figure 1, mcMPO-Gd differs from the previously reported compound "MPO-Gd" in that the two 5-hydroxyindole moieties (5-hydroxytryptophan and 5-hydroxyindoleacetic acid) are offset and distanced from one side of the chelate (rather than from the opposite side of the chelate) by a short linker. The two amide bonds formed by the linker of mcMPO-Gd increase the rigidity of the agent compared to MPO-Gd. Without being bound by theory, it is hypothesized that the rigid structure of mcMPO-Gd would produce a more potent agent than MPO-Gd by 1) reducing segment motion to increase relaxation potency and 2) creating better protein binding efficiency to increase sensitivity to MPO activity.

The reactions used to prepare the compounds and salts described herein can be carried out in a suitable solvent readily selected by those skilled in the art of organic synthesis. Suitable solvents can be substantially non-reactive with the starting materials (reactants), intermediates, or products at the temperatures at which the reaction is carried out (e.g., temperatures that can range from the freezing temperature of the solvent to the boiling temperature of the solvent). A given reaction may be carried out in one solvent or a mixture of more than one solvent. Depending on the particular reaction step, a suitable solvent for the particular reaction step may be selected by the skilled person.

The preparation of the compounds and salts described herein may involve the protection and deprotection of various chemical groups. The need for protection and deprotection and the choice of an appropriate protecting group can be readily determined by those skilled in the art. The chemical nature of the protecting Groups can be found, for example, in T.W.Greene and P.G.M.Wuts, "Protective Groups in Organic Synthesis", 3 rd edition, Wiley & Sons, Inc., New York, Inc. (1999).

The reaction may be monitored according to any suitable method known in the art. For example, the magnetic resonance spectrum can be obtained by, for example, ¹ h or ¹³ C) By spectroscopic means such as infrared spectroscopy, spectrophotometry (e.g., UV visible), mass spectrometry, or by methods such as High Performance Liquid Chromatography (HPLC),Liquid chromatography-mass spectrometry (LCMS) or Thin Layer Chromatography (TLC) to monitor product formation. Purification of the compounds can be carried out by various methods by those skilled in the art, including High Performance Liquid Chromatography (HPLC) and normal phase silica gel chromatography.

In various positions in this specification, divalent linking substituents are described. It is particularly desirable that each divalent linking substituent comprises both the forward and reverse forms of the linking substituent. For example, -NR (CR 'R') _n -comprises-NR (CR 'R') _n -and- (CR 'R') _n NR-both; and NHC (O) L ² comprising-NHC (O) L ² -and-L ² C (O) NH-. In the case of structures in which a linking group is explicitly required, the markush variables (markush variable) listed in connection with the group are to be understood as linking groups.

The term "n-membered" (where n is an integer) generally describes the number of ring-forming atoms in the moiety, where the number of ring-forming atoms is n. For example, phenyl is an example of a 6-membered aryl ring, pyrazolyl is an example of a 5-membered heteroaryl ring, and pyridyl is an example of a 6-membered heteroaryl ring.

Throughout the definition, the term "C _n-m "indicates ranges including endpoints where n and m are integers and indicate a number of carbons. Examples include C _1-4 、C _1-6 And so on.

As used herein, the term "C _n-m Alkyl "refers to a saturated hydrocarbon group having n to m carbons that may be straight or branched. Examples of alkyl moieties include, but are not limited to, chemical groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, and the like. In some embodiments, the alkyl group contains 1 to 6 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms.

As used herein, the term "C _n-m Alkylene "refers to a divalent alkyl linking group having n to m carbons (e.g., -CH) ₂ -, eth-1, 2-diyl, prop-1, 3-diyl, etc.). In some embodiments, the alkylene moiety contains 2 to 6,2 to 4, 2 to 3,1 to 6,1 to 4, or 1 to 2 carbon atoms.

As herein describedAs used, the term "C _n-m Alkenylene "refers to a divalent olefinic linking group having n to m carbons. In some embodiments, the alkenylene moiety contains 2 to 6,2 to 4, or 2 to 3 carbon atoms.

As used herein, the term "C _n-m Alkyleneoxy "refers to a divalent alkoxy linking group having n to m carbons (i.e.," -O-C) _n-m Alkylene- "). In some embodiments, the alkyleneoxy moiety contains 1 to 2 to 6,2 to 4, 2 to 3,1 to 6,1 to 4, or 1 to 2 carbon atoms.

As used herein, the term "aryl" refers to an aromatic hydrocarbon group that may be monocyclic or polycyclic (e.g., having 2, 3, or 4 fused rings). The term "C _n-m Aryl "refers to an aryl group having n to m ring carbon atoms. Aryl groups include, for example, phenyl, naphthyl, anthryl, phenanthryl, indanyl, indenyl, and the like. In some embodiments, the aryl group has 6 to about 20 carbon atoms, 6 to about 15 carbon atoms, or 6 to about 10 carbon atoms. In some embodiments, aryl is phenyl.

As used herein, "heteroaryl" refers to a monocyclic or polycyclic aromatic heterocycle having at least one heteroatom ring member selected from sulfur, oxygen, and nitrogen. In some embodiments, the heteroaryl ring has 1,2, 3, or 4 heteroatom ring members independently selected from nitrogen, sulfur, and oxygen. In some embodiments, any ring-forming N in the heteroaryl moiety can be an N-oxide. In some embodiments, heteroaryl has 5-20 ring atoms and 1,2, 3, or 4 heteroatom ring members independently selected from nitrogen, sulfur, and oxygen. In some embodiments, heteroaryl has 5-16 ring atoms and 1,2, 3, or 4 heteroatom ring members independently selected from nitrogen, sulfur, and oxygen. In some embodiments, the heteroaryl is a bicyclic heteroaryl (e.g., fused bicyclic heteroaryl) having 5 to 16 ring atoms and 1,2, 3, or 4 heteroatom ring members independently selected from nitrogen, sulfur, and oxygen. In some embodiments, heteroaryl is a tricyclic heteroaryl (e.g., fused tricyclic heteroaryl) having 5 to 16 ring atoms and 1,2, 3, or 4 heteroatom ring members independently selected from nitrogen, sulfur, and oxygen.

As used herein, "heteroCycloalkyl "refers to a non-aromatic monocyclic or polycyclic heterocycle having one or more ring-forming heteroatoms selected from O, N or S. The ring-forming carbon atoms and heteroatoms of the heterocycloalkyl group may be optionally substituted with an oxylene or sulfide group (e.g., C (═ O), S (═ O), C (S), or S (═ O) ₂ Etc.). The heterocycloalkyl group may be attached through a ring-forming carbon atom or a ring-forming heteroatom. In some embodiments, the heterocycloalkyl group contains 0 to 3 double bonds or 0 to 2 double bonds. The heterocycloalkyl group containing a fused aromatic ring may be linked through any ring-forming atom including the ring-forming atoms of the fused aromatic ring. In some embodiments, heterocycloalkyl has 4-20, 4-16, 8-20, or 8-16 ring atoms with 1 or 2 heteroatoms independently selected from nitrogen, oxygen, or sulfur and optionally one or more oxygenated ring members. In some embodiments, the heterocycloalkyl group is a bicyclic heterocycloalkyl group (e.g., a fused bicyclic heterocycloalkyl group) having 4-16 ring atoms and 1,2, 3, or 4 heteroatom ring members independently selected from nitrogen, sulfur, and oxygen, and optionally having one or more oxygenated ring members. In some embodiments, the heterocycloalkyl group is a tricyclic heterocycloalkyl group (e.g., a fused tricyclic heterocycloalkyl group) having 4 to 16 ring atoms and 1,2, 3, or 4 heteroatom ring members independently selected from nitrogen, sulfur, and oxygen, and optionally having one or more oxygenated ring members.

In certain places, definitions or embodiments refer to particular rings (e.g., azetidine rings, pyridine rings, etc.). Unless otherwise indicated, these rings may be attached to any ring member, provided that the valency of the atoms is not exceeded. For example, the pyridine ring may be attached at any position on the ring, while the pyridine-3-yl ring is attached at the 3-position.

As used herein, the term "compound" is intended to encompass all stereoisomers, geometric isomers, tautomers, and isotopes of the depicted structures. Unless otherwise specified, a compound identified herein by name or structure as one particular tautomeric form is intended to encompass other tautomeric forms.

The compounds provided herein also include tautomeric forms. The tautomeric forms result from the exchange of a single bond with an adjacent double bond and the concomitant migration of protons. Tautomeric forms include prototropic tautomers, which are isomeric protonation states having the same empirical formula and total charge. Exemplary prototropic tautomers include keto-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, enamine-imine pairs and cyclic forms, wherein protons may occupy two or more positions of a heterocyclic ring system, such as 1H-and 3H-imidazole, 1H-, 2H-and 4H-1,2, 4-triazole, 1H-and 2H-isoindole and 1H-and 2H-pyrazole. Tautomeric forms can be in equilibrium or, by appropriate substitution, sterically locked into one form.

Unless otherwise specifically stated, the compounds provided herein may also contain all of the atomic isotopes present in the intermediates or final compounds. Isotopes include those atoms having the same atomic number but different mass numbers. Unless otherwise specified, when an atom is designated as an isotope or radioisotope, the atom is understood to include the isotope or radioisotope in an amount at least greater than the natural abundance of the isotope or radioisotope. For example, when an atom is designated as "D" or "deuterium", the position is understood to be deuterium having an abundance that is at least 3000 times greater than the natural abundance of deuterium, which is 0.015% (i.e., at least 45% deuterium incorporation).

All compounds and their pharmaceutically acceptable salts may be found together with other substances such as water and solvents (e.g. hydrates and solvates) or may be isolated.

In some embodiments, preparation of the compounds may involve the addition of an acid or base to affect, for example, catalysis of the desired reaction or formation of salt forms such as acid addition salts.

Exemplary acids may be inorganic or organic acids and include, but are not limited to, strong and weak acids. Some exemplary acids include hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, p-toluenesulfonic acid, 4-nitrobenzoic acid, methanesulfonic acid, benzenesulfonic acid, trifluoroacetic acid, and nitric acid. Some weak acids include, but are not limited to, acetic acid, propionic acid, butyric acid, benzoic acid, tartaric acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, and capric acid.

Exemplary bases include lithium hydroxide, sodium hydroxide, potassium hydroxide, lithium carbonate, sodium carbonate, potassium carbonate, and sodium bicarbonate. Some exemplary strong bases include, but are not limited to, hydroxides, alkoxides, metal amides, metal hydrides, metal dialkylamides, and arylamines, wherein alkoxides include lithium, sodium, and potassium salts of methyl, ethyl, and tert-butyl oxides; the metal amide comprises sodium amide, potassium amide and lithium amide; the metal hydride includes sodium hydride, potassium hydride, and lithium hydride; and the metal dialkylamides comprise lithium, sodium and potassium salts of methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, trimethylsilyl and cyclohexyl substituted amides.

In some embodiments, a compound provided herein or a salt thereof is substantially isolated. By "substantially isolated" is meant that the compound is at least partially or substantially separated from the environment in which it is formed or detected. Partial isolation may comprise, for example, compositions enriched in the compounds provided herein. Substantially isolating may comprise a composition containing at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% by weight of a compound provided herein, or a salt thereof. Methods for isolating compounds and salts thereof are conventional in the art.

As used herein, the term "room temperature" or "rt" is understood in the art and generally refers to a temperature, such as a reaction temperature, i.e., the approximate temperature of the room in which the reaction is conducted, e.g., a temperature of from about 20 ℃ to about 30 ℃.

The phrase "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

The present application also encompasses pharmaceutically acceptable salts of the compounds described herein. As used herein, "pharmaceutically acceptable salts" refer to derivatives of the disclosed compounds wherein the parent compound is modified by conversion of an existing acid or base moiety into its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines; acidic residues such as alkali or organic salts of carboxylic acids, and the like. The pharmaceutically acceptable salts herein include the conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of the present application can be synthesized from the parent compound, which contains a basic or acidic moiety, by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or a mixture of the two; generally, nonaqueous media such as diethyl ether, ethyl acetate, alcohols (e.g. methanol, ethanol, isopropanol or butanol) or acetonitrile (MeCN) are preferred. A list of suitable salts is presented in Remington's Pharmaceutical Sciences, 17 th edition, Mark Press, Isston, Pa., 1985, p. 1418 and Journal of Pharmaceutical Sciences, 66,2 (1977). For example, in the handbook of pharmaceutical salts: properties, Selection and uses (Handbook of Pharmaceutical Salts, Selection, and Use), Wiley-VCH,2002, describe a conventional process for preparing salt forms.

Application method

The present application further provides a method of imaging a cell or tissue sample. As used herein, the term "subject" refers to any animal, including mammals. For example, mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, primates, and humans. In some embodiments, the subject is a human.

In some embodiments, the method comprises:

i) administering to a subject a compound provided herein (e.g., a compound of any one of formulas I-Vc, or a pharmaceutically acceptable salt thereof);

iii) imaging the cell or tissue sample using an imaging technique. In some embodiments, the method further comprises imaging the cell or tissue sample prior to step i). In some embodiments, the method is an in vitro method. In some embodiments, the method is an in vivo method.

The present application further provides a method of diagnosing a disease or disorder associated with aberrant Myeloperoxidase (MPO) activity in a subject. In some embodiments, the method comprises:

i) administering to the subject a compound provided herein (e.g., a compound of any one of formulas I-Vc, or a pharmaceutically acceptable salt thereof);

iii) imaging the cell or tissue using imaging techniques. In some embodiments, the method further comprises imaging the subject prior to step i). In some embodiments, the method is an in vitro method. In some embodiments, the method is an in vivo method.

In some embodiments, sufficient time is from about 5 minutes to about 6 hours, such as from about 5 minutes to about 6 hours, from about 5 minutes to about 4 hours, from about 5 minutes to about 2 hours, from about 5 minutes to about 1 hour, from about 5 minutes to about 30 minutes, from about 30 minutes to about 6 hours, from about 30 minutes to about 4 hours, from about 30 minutes to about 2 hours, from about 30 minutes to about 1 hour, from about 1 hour to about 6 hours, from about 1 hour to about 4 hours, from about 1 hour to about 2 hours, from about 2 hours to about 6 hours, from about 2 hours to about 4 hours, or from about 4 hours to about 6 hours.

The present application further provides a method of imaging Myeloperoxidase (MPO) activity in a cell. In some embodiments, the method comprises:

i) contacting the cell with a compound provided herein (e.g., a compound of any one of formulas I-Vc, or a pharmaceutically acceptable salt thereof); and

ii) imaging the cell with an imaging technique.

The present application further provides a method of imaging Myeloperoxidase (MPO) activity in a tissue sample. In some embodiments, the method comprises:

i) contacting the tissue sample with a compound provided herein (e.g., a compound of any one of formulas I-Vc, or a pharmaceutically acceptable salt thereof); and

ii) imaging the tissue sample with an imaging technique.

The present application further provides a method of detecting Myeloperoxidase (MPO) activity in a cell or tissue sample. In some embodiments, the method comprises:

i) contacting the cell or tissue sample with a compound provided herein (e.g., a compound of any one of formulas I-Vc, or a pharmaceutically acceptable salt thereof); and

ii) imaging the cell or tissue sample with an imaging technique.

The present application further provides a method of detecting myeloperoxidase activity in a subject. In some embodiments, the method comprises:

i) administering to the subject a compound provided herein (e.g., a compound of any one of formulas I-Vc, or a pharmaceutically acceptable salt thereof); and

ii) imaging the subject with an imaging technique.

The present application further provides a method of monitoring treatment of a disease or disorder associated with abnormal Myeloperoxidase (MPO) activity in a subject, the method comprising:

ii) imaging the subject with an imaging technique;

iv) imaging the cells or tissues of the subject with an imaging technique; and

v) comparing the image of step i) with the image of step iv).

In some embodiments, the method further comprises administering to the subject a compound provided herein (e.g., a compound of any one of formulas I-Vc, or a pharmaceutically acceptable salt thereof) after the administering of step iii) and before the imaging of step iv). In some embodiments, the therapeutic compounds are useful for treating diseases or disorders associated with abnormal Myeloperoxidase (MPO) activity. In some embodiments, the therapeutic compound is a therapeutic compound provided herein.

In some embodiments, the imaging technique is selected from the group consisting of magnetic resonance imaging and nuclear imaging. In some embodiments, the imaging technique is magnetic resonance imaging. In some embodiments, the imaging technique is nuclear imaging.

In some embodiments, the compound of formula I is:

or a pharmaceutically acceptable salt thereof, and the imaging technique is selected from the group consisting of magnetic resonance imaging and nuclear imaging. In some embodiments, the imaging technique is magnetic resonance imaging. In some embodiments, the imaging technique is nuclear imaging.

In some embodiments, the disease or disorder associated with abnormal myeloperoxidase activity is selected from the group consisting of: non-alcoholic steatohepatitis (NASH), cancer, rheumatic diseases, infectious diseases, central nervous system diseases, cardiovascular disorders, autoimmune disorders, and inflammation associated with one or more of cancer, rheumatic diseases, infectious diseases, central nervous system diseases, cardiovascular disorders, and autoimmune disorders. In some embodiments, the disease or disorder associated with abnormal myeloperoxidase activity is selected from the group consisting of: cancer, rheumatic diseases, infectious diseases, central nervous system diseases, cardiovascular disorders and autoimmune disorders. In some embodiments, the disease or disorder associated with abnormal myeloperoxidase activity is selected from the group consisting of: inflammation associated with one or more of cancer, rheumatic diseases, infectious diseases, central nervous system diseases, cardiovascular disorders, and autoimmune disorders.

In some embodiments, the disease or disorder associated with aberrant myeloperoxidase activity is cancer. In some embodiments, the cancer comprises a solid tumor. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is selected from the group consisting of: bladder cancer, breast cancer, carcinoma, cervical cancer, colorectal cancer, endometrial cancer, glioma, head and neck cancer, liver cancer, lung cancer, melanoma, ovarian cancer, pancreatic cancer, prostate cancer, renal cancer, skin cancer, stomach cancer, testicular cancer, leukemia, and thyroid cancer. In some embodiments, the cancer is a solid tumor associated with one or more of bladder cancer, breast cancer, carcinoma, cervical cancer, colorectal cancer, endometrial cancer, glioma, head and neck cancer, liver cancer, lung cancer, melanoma, ovarian cancer, pancreatic cancer, prostate cancer, renal cancer, skin cancer, gastric cancer, testicular cancer, thyroid cancer, or any combination thereof. In some embodiments, the disease or disorder associated with aberrant myeloperoxidase activity is inflammation associated with one or more cancers selected from the group consisting of: bladder cancer, breast cancer, carcinoma, cervical cancer, colorectal cancer, endometrial cancer, glioma, head and neck cancer, liver cancer, lung cancer, melanoma, ovarian cancer, pancreatic cancer, prostate cancer, renal cancer, skin cancer, stomach cancer, testicular cancer, thyroid cancer, leukemia, or any combination thereof.

In some embodiments, the disease or disorder associated with abnormal myeloperoxidase activity is a central nervous system disease. In some embodiments, the central nervous system disease is selected from the group consisting of: alzheimer's disease, stroke, epilepsy, Parkinson's disease, and inflammation associated with Alzheimer's disease, stroke, epilepsy, and Parkinson's disease. In some embodiments, the central nervous system disease is selected from the group consisting of: alzheimer's disease, stroke, epilepsy, and parkinson's disease. In some embodiments, the central nervous system disorder is inflammation associated with one or more of alzheimer's disease, stroke, epilepsy, and parkinson's disease.

In some embodiments, the disease or disorder associated with aberrant myeloperoxidase activity is a cardiovascular disorder. In some embodiments, the cardiovascular disorder is selected from the group consisting of: atherosclerosis, myocardial infarction, atrial fibrillation, vasculitis, and inflammation associated with one or more of atherosclerosis, myocardial infarction, atrial fibrillation, and vasculitis. In some embodiments, the cardiovascular disorder is selected from the group consisting of atherosclerosis, myocardial infarction, atrial fibrillation, and vasculitis. In some embodiments, the cardiovascular disorder is inflammation associated with one or more of atherosclerosis, myocardial infarction, atrial fibrillation, and vasculitis.

In some embodiments, the disease or disorder associated with aberrant myeloperoxidase activity is an autoimmune disorder. In some embodiments, the autoimmune disorder is selected from the group consisting of: multiple sclerosis, meningitis, encephalitis, and inflammation associated with one or more of multiple sclerosis, meningitis, and encephalitis. In some embodiments, the autoimmune disorder is inflammation associated with one or more of multiple sclerosis, meningitis, and encephalitis.

In some embodiments, the disease or disorder associated with aberrant myeloperoxidase activity is a rheumatic disease. In some embodiments, the rheumatic disease is selected from the group consisting of rheumatoid arthritis, osteoarthritis, and inflammatory arthritis. In some embodiments, the rheumatic disease is inflammatory arthritis. In some embodiments, the inflammatory arthritis is selected from the group consisting of gout and calcium pyrophosphate deposition disease (CPPD). In some embodiments, the disease or disorder associated with aberrant myeloperoxidase activity is inflammation associated with one or more of rheumatoid arthritis, osteoarthritis, and inflammatory arthritis.

In some embodiments, the disease or disorder associated with abnormal myeloperoxidase activity is an infectious disease. In some embodiments, the infectious disease is a fungal disease or a bacterial disease. In some embodiments, the fungal disease is a disease associated with candida albicans (c. In some embodiments, the infectious disease comprises a yeast infection. In some embodiments, the yeast infection is an infection associated with candida tropicalis (c. In some embodiments, the disease or disorder associated with abnormal myeloperoxidase activity is inflammation associated with an infectious disease or bacterial disease.

As used herein, the phrase "therapeutically effective amount" refers to the amount of active compound or pharmaceutical agent that elicits the biological or medicinal response that a researcher, veterinarian, medical doctor or other clinician seeks to in a tissue, system, animal, individual, or human. In some embodiments, the dose of the compound or pharmaceutically acceptable salt thereof administered to the subject or individual is from about 1mg to about 2g, from about 1mg to about 1000mg, from about 1mg to about 500mg, from about 1mg to about 100mg, from about 1mg to 50mg, or from about 50mg to about 500 mg.

As used herein, the term "treating" or "treatment" refers to one or more of the following: (1) inhibiting the disease; for example, inhibiting a disease, condition, or disorder in an individual who is experiencing or exhibiting a pathology or symptomatology of the disease, condition, or disorder (i.e., arresting further development of the pathology and/or symptomatology); and 2) ameliorating the disease; for example, ameliorating a disease, condition, or disorder (i.e., reversing the pathology and/or symptomatology) in an individual who is experiencing or exhibiting the pathology or symptomatology of the disease, condition, or disorder, such as reducing the severity of the disease or alleviating or relieving one or more symptoms of the disease.

Combination therapy

One or more additional therapeutic agents, e.g., anti-inflammatory agents, steroids, immunosuppressive agents, chemotherapeutic agents, or other agents such as therapeutic antibodies may be used in combination with the compounds and salts of the present application for the treatment of the diseases provided herein.

Exemplary antibodies for use in combination therapy include, but are not limited to, trastuzumab (trastuzumab) (e.g., anti-HER 2), ranibizumab (ranibizumab) (e.g., anti-VEGF-a), bevacizumab (e.g., anti-VEGF), panitumumab (e.g., anti-EGFR), cetuximab (cetuximab) (e.g., anti-EGFR), rituximab (rituxan) (anti-CD 20), and antibodies against c-MET.

Exemplary steroids include corticosteroids such as cortisone (cortisone), dexamethasone (dexamethasone), hydrocortisone (hydrocortisone), methylprednisolone (methylprednisone), prednisolone (prednisone), and prednisone (prednisone).

Exemplary anti-inflammatory compounds include aspirin (aspirin), choline salicylate, celecoxib (celecoxib), diclofenac potassium (diclofenac potassium), diclofenac sodium and misoprostol (misoprostol), diflunisal (diflunisa), etodolac (etodolac), fenoprofen (fenoprofen), flurbiprofen (flurbiprofen), ibuprofen (uprofen), ketoprofen (ketoprofen), meclofenamate sodium (meclofenamate sodium), mefenamic acid (mefenamic acid), nabumetone (nabumetone), naproxen (naproxen), naproxen sodium (naproxen sodium), oxaprozin (oxaprozin), piroxicam (piroxicam), rofecoxib (salfenac), salbutate (salsulindac), and sodium docusate (salbutamide).

Exemplary immunosuppressive agents include azathioprine (azathioprine), chlorambucil (chlorambucil), cyclophosphamide (cyclophosphamide), cyclosporine (cyclosporine), daclizumab (daclizumab), infliximab (infliximab), methotrexate (methotrexate), and tacrolimus (tacrolimus).

One or more of the following agents may be used in combination with the compounds provided herein and presented as a non-limiting list: cell growth inhibitors, cisplatin (cispain), doxorubicin (doxorubicin), taxol (taxol), etoposide (etoposide), irinotecan (irinotecan), topotecan (topotecan), paclitaxel (paclitaxel), docetaxel (docetaxel), epothilones (epothilones), tamoxifen (tamoxifen), 5-fluorouracil, methotrexate, temozolomide (temozolomide), cyclophosphamide, tipifarnib (tipifarnib), gefitinib (gefitinib), erlotinib hydrochloride (erlotinib hydrochloride), EGFR antibodies, imatinib mesylate (imatinib mesylate), gemcitabine (gemcitabine), uramustine (uracil), nitrogen mustard (loximine), ifosfamide (amiferamide), melphalan (melamine), melamine (sulfacetamide), melphalan (bupropion (ethyl-e), docetaxel (docetaxel), docetaxel (docetaxel), a (docetaxel (a), a pharmaceutically acceptable salt (e), a pharmaceutically acceptable salt (a pharmaceutically acceptable salt, a, Streptozotocin (streptozocin), dacarbazine (dacarbazine), floxuridine (floxuridine), cytarabine (cytarabine), 6-mercaptopurine, 6-thioguanine (thioguanine), fludarabine phosphate (fludarabine phosphate), oxaliplatin (oxaliplatin), folinic acid, pentostatin (pentostatin), vinblastine (vinblastine), vincristine (vincristine), vindesine (vindesine), bleomycin (bleomycin), actinomycin (dactinomycin), daunorubicin (daunorubicin), epirubicin (epirubicin), idarubicin (idarubicin), mithramycin (mithramycin), mithramycin (medrypsin), doxycycline (doxycycline), mitomycin-C (mitomycin-C), L-asparaginase (L-asparaginase), nisin (17-ketoprofen), prednisolone (alpha-17-ketoprofen), dihydrocodeine (diethylstilben), dihydrocodeine (dihydrocodeine), dihydrocodeine (dihydrocodeine, dihydrocodeine (dihydrocodeine), dihydrocodeine (dihydrocodeine, beta-C, beta-D-, Testolactone (testolactone), megestrol acetate (megestrol acetate), methylprednisolone, methyltestosterone (methyltestosterone), prednisolone, triamcinolone (triamcinolone), chlorfenapyr (chlorotrianisene), hydroxyprogesterone (hydroxyprogesterone), aminoglutethimide (aminoglutethimide), estramustine (estramustine), medroxyprogesterone acetate (medroxyprogesterone), leuproreline (leurolide), flutamide (flutamide), toremifene (toremifene), goserelin (serelin), carboplatin, hydroxyurea (hydroxyurea), amsacrine (amsacrine), procarbazine (procarbazine), mitotane (mitoxantrone), mitoxantrone (mitoxantrone), meglumine (vamidoxime), levofloxacin (levofloxacin), levofloxacin (valtrexadine), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin (levofloxacin), levofloxacin, Erbitux, thiotepa (thiotepa), altretamine (altretamine), trastuzumab (trastuzumab), fluvastatin (fulvestrant), exemestane (exemestane), rituximab (rituximab), alemtuzumab (alemtuzumab), clofarabine (clofarabine), cladribine (cladribine), aphidicolin (aphidicolin), sunitinib (sunitinib), dasatinib (dasatinib), tizacitabine (tezacitabine), triazapine (triapin), didodecine (didox), trimazole (trimido), amimetx (amidox), bendamustine (bendamusine), orimazumab (ofatumumab), and elvisib (elvisib).

In some embodiments, the additional therapeutic agent may be used to treat multiple sclerosis. In some embodiments, the additional therapeutic agent is selected from the group consisting of: interferon beta-1 a, interferon beta-1 b, pegylated interferon beta-1 a, glatiramer acetate, teriflunomide (teriflunomide), fingolimod, mitoxantrone, dimethyl fumarate, natalizumab (natalizumab), ozagrimod (ozanimod), laquinimod (laquinimod), alemtuzumab, darlizumab, rituximab, ocrelizumab (ocrelizumab), and ofatumumab.

Pharmaceutical formulation

When used as a medicament, the compounds and salts provided herein may be administered in the form of a pharmaceutical composition. These compositions may be prepared as described herein or elsewhere and may be administered by a variety of routes depending on whether local or systemic treatment is desired and the area to be treated. Administration can be topical (including transdermal, epidermal, ocular and to mucous membranes, including intranasal, vaginal and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal or intranasal), oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal intramuscular or injection or infusion; or intracranial (e.g., intrathecal or intracerebroventricular administration). Parenteral administration may be in the form of a single bolus dose, or may be performed, for example, by a continuous infusion pump. In some embodiments, the compounds provided herein are suitable for parenteral administration. In some embodiments, the compounds provided herein are suitable for intravenous administration. Pharmaceutical compositions and formulations for topical administration may comprise transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable. In some embodiments, the pharmaceutical compositions provided herein are suitable for parenteral administration. In some embodiments, the compositions provided herein are suitable for intravenous administration.

Also provided are pharmaceutical compositions containing a compound provided herein as an active ingredient (e.g., a compound of any one of formulas I-Vc, or a pharmaceutically acceptable salt thereof) and one or more pharmaceutically acceptable carriers (excipients). In preparing the compositions provided herein, the active ingredient is typically mixed with, diluted with, or encapsulated within an excipient such carrier, e.g., in the form of a capsule, sachet (sachet), paper, or other container. When the excipient serves as a diluent, it may be a solid, semi-solid, or liquid material that can act as a vehicle, carrier, or medium for the active ingredient. Thus, the compositions may be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.

Some examples of suitable excipients include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methyl cellulose. The formulation may also include, but is not limited to, lubricants such as talc, magnesium stearate, and mineral oil; a wetting agent; emulsifying and suspending agents; preservatives, such as methyl and propyl hydroxybenzoate; a sweetener; a flavoring agent, or a combination thereof.

The active compounds may be effective over a wide dosage range and are generally administered in a pharmaceutically effective amount. It will be understood, however, that the amount of the compound actually administered will generally be determined by a physician, in the light of the relevant circumstances, including the chosen route of administration, the actual compound administered, the age, weight, and response of the individual subject, the severity of the subject's symptoms, and the like.

Examples of the invention

The present invention will be described in more detail by specific examples. The following examples are provided for illustrative purposes and are not intended to limit the invention in any way.

General methods and materials

All chemicals were obtained from Sigma Chemical company (Sigma Chemical Co.) unless otherwise noted. 5-hydroxy-L-tryptophan was obtained from Chem-Impex International Inc. (Chem-Impex International Inc.) (Wood Dale, IL.). DO 3A-tert-butyl ester was obtained from Macrocyclics Inc. (Plano, TX)), and serotonin was obtained from TCI America (TCI America) (Portland, Oregon). Glucose oxidase was obtained from alfenmeretrix corporation (Affymetrix) (Santa Clara, CA). Complete Freund's Adjuvant was purchased from Sigma-Aldrich (Sigma-Aldrich). Myeloperoxidase is available from Lee Biosolutions, st. Using Thermo Scientific ^TM Q-active Plus Ultimate3000 HPLC flow injection analysis for high resolution mass spectrometry. Using Bruker Ascend ^TM 400 proceed with ¹ H-NMR and ¹³ C-NMR. Liquid chromatography mass spectrometry was performed using a Waters 2545 binary gradient module with a 2777 sample manager.

The normality of all digital data was first analyzed using the Shapiro-Wilk normality test (Shapiro-Wilk normative test), followed by determination of significance using the appropriate parametric or non-parametric test. Evaluation of TS Apoe Using the Kruskal-Wallis test (Kruskal-Wallis test) and Dunn multiple comparison (Dunn's multiple compare), Mantney rank sum test (Mann-Whitney rank sum test) or, where appropriate, unpaired t test ^–/– And Mpo ^–/– Apoe ^–/– TS Apoe between or fed WD + -AZM 198 ^–/– Δ CNR difference between. All statistical analyses were performed using GraphPad Prism version 8.01 (GraphPad Software, La Jolla California, ca, USA) adapted for Mac, and individual data are shown as mean ± SEM. P value<0.05 was considered significant.

Example 1: synthesis of di-tert-butyl (S) -2-bromoglutarate (Compound 1)

Compound 1 was prepared according to the previously reported procedure (see, e.g., International patent application No. WO 2005/122682; and Moumne et al, J. org. chem. 2006,71: 3332-3334).

Step 1: (S) -2-Bromoglutaric acid

Adding NaNO ₂ A solution of (0.9g, 13mmol) in water (5mL) was added dropwise to a mixture of L-glutamic acid (1.47g, 10mmol) and NaBr (3.8g, 37mmol) in 0.75M HBr (30mL) at-15 deg.C over 30 minutes. The reaction was stirred at the same temperature for an additional 2 hours, then concentrated H was slowly added to the solution ₂ SO ₄ (1mL) then Et ₂ O (30 mL. times.3). The combined organic phases were washed with brine, over Na ₂ SO ₄ Dried and evaporated to give (S) -2-bromoglutaric acid, which is used without further purificationAnd (4) carrying out the next step.

Step 2: (S) -2-Bromoglutaric acid di-tert-butyl ester (Compound 1)

MgSO (MgSO) ₄ (5g) And concentrated H ₂ SO ₄ A mixture (0.5mL) in DCM (5mL) was stirred at room temperature for 2h, then the above compound and t-BuOH (3g, 40mmol) were added and stirred for another 20 h. The reaction was filtered to remove salts and extracted with DCM (20mL × 3). The combined organic layers were washed with brine, over Na ₂ SO ₄ Dried, evaporated and subjected to flash chromatography (hexane: ethyl acetate 5:1) to give compound 1 as a yellow oil (45%, for two steps). ¹ H NMR(500MHz,CDCl ₃ )δ4.23(dd,1H),2.51(m,2H),2.29(m,2H),1.48(s,9H),1.43(s,9H)； ¹³ C NMR(125MHz,CDCl ₃ ) δ 172.7,168.4,82.8,82.6,46.8,32.9,30.0,28.1, 27.9. LCMS found m/z (ES +): 325.3(M + H).

Example 2: synthesis of (S) -2-amino-3- (5-hydroxy-1H-indol-3-yl) -N- (2- (5-hydroxy-1H-indol-3-yl) ethyl) propionamide (intermediate 4)

Step 1: (S) -2- ((tert-butoxycarbonyl) amino) -3- (5-hydroxy-1H-indol-3-yl) propionic acid (Compound 2)

A solution of di-tert-butyl dicarbonate (784mg, 3.6mmol) in THF (4mL) was added to L-5-hydroxy-tryptophan (5-HTP, 660mg, 3.0mmol) and K ₂ CO ₃ (880mg, 6.4mmol) in water (8 mL). The reaction was stirred at room temperature for 2 hours and then neutralized to pH 2-3 by addition of 1M HCl. After evaporation to remove THF, the solution was extracted with ethyl acetate (20 mL. times.3)The organic phase was washed with brine (10 mL. times.3) and dried over anhydrous Na ₂ CO ₃ Dried and evaporated to give compound 2, which was used in the next step without further purification.

Step 2: (S) - (3- (5-hydroxy-1H-indol-3-yl) -1- ((2- (5-hydroxy-1H-indol-3-yl) ethyl) amino) -1-oxopropan-2-yl) carbamic acid tert-butyl ester (Compound 3)

To a solution of compound 2(385mg, 1.2mmol) in DMF (5mL) was added edc.hcl (280mg, 1.4mmol) followed by HOBt (216mg, 1.4mmol) and the resulting mixture was stirred for 10 min. Next, a solution of free serotonin (220mg, 1.0mmol) prepared in advance in DMF (4mL) was added to the reaction mixture. The resulting mixture was then stirred for an additional 2 hours. The reaction mixture was extracted with ethyl acetate (10 mL. times.3), and the organic layer was washed with brine (10 mL. times.3) and dried over anhydrous Na ₂ CO ₃ Dried and evaporated. The residue was purified by flash chromatography (ethyl acetate as eluent) to give compound 3(390mg) as a white solid in 82% yield. ¹ H NMR(500MHz,DMSO)δ10.45(s,2H),8.56(s,1H),8.54(s,1H),7.89(t,1H),7.10(d,1H),7.09(d,1H),7.0(m,2H),6.88(d,1H),6.83(d,1H),6.67(d,1H),6.57(m,2H),4.12(m,1H),3.26(m,2H),2.96(dd,1H),2.79(dd,1H),2.66(m,2H),1.32(s,9H)； ¹³ C NMR (125MHz, DMSO) δ 171.8,155.1,150.15,150.13,130.8,130.6,128.1,127.8,123.9,123.0,111.6,111.4,111.2,111.1,110.7,109.3,102.5,102.2,77.9,55.0,40.1,28.2,28.0, 25.2; LCMS found m/z: 479.4(M + H).

And step 3: (S) -2-amino-3- (5-hydroxy-1H-indol-3-yl) -N- (2- (5-hydroxy-1H-indol-3-yl) ethyl) propionamide (intermediate 4)

Compound 3(300mg) was added to 10% trifluoroacetic acid(TFA) in dichloromethane (DCM; 4mL) and the reaction mixture was stirred at room temperature for 5 h. The reaction mixture was then evaporated under reduced pressure to remove the solvent and subjected to preparative HPLC to give the desired intermediate 4 (63% yield). ¹ H NMR(500MHz,DMSO)δ10.70(d,1H),10.50(d,1H),8.57(m,3H),8.05(m,2H),7.14(dd,2H),7.09(d,1H),6.98(dd,2H),6.81(d,1H),6.62(dd,2H),3.87(dt,1H),3.34(m,2H),3.10(dd,1H),2.98(dd,1H),2.68(m,2H)； ¹³ C NMR (125MHz, DMSO). delta. 168.3,150.4,150.2,130.9,130.8,127.8,127.7,125.2,123.2,111.7,111.5,111.2,110.4,106.0,102.6,102.1,94.8,52.8,39.9,27.6, 25.0. LCMS found m/z: 379.5(M + H).

Example 3: synthesis of di-tert-butyl (R) -2- (4,7, 10-tris (2- (tert-butoxy) -2-oxoethyl) -1,4,7, 10-tetraazacyclododecan-1-yl) glutarate (Compound 6)

Potassium carbonate (210mg, 1.5mmol) was added to tri-tert-butyl 2,2' - (1,4,7, 10-tetraazacyclododecane-1, 4, 7-triyl) triacetate (DO) ₃ A-tBu-ester; compound 5; 770mg, 1.5mmol) and compound 1(480mg, 1.5mmol) in acetonitrile (5mL) and the reaction mixture was heated at reflux for 24 h. After removal of the solvent under reduced pressure, the residue was dissolved in DCM and filtered. Flash column eluting with a gradient of DCM to DCM containing 5% MeOH afforded compound 6 as a light yellow solid in 78% yield. ¹ H NMR(500MHz,DMSO)δ3.48-1.90(m,27H),1.40(s,45H) ¹³ C NMR (125MHz, DMSO)174.8,173.1,173.0,172.3,82.5,82.0,81.9,80.6,60.0,55.9(2),55.6,52.7,52.5,48.6,48.1,47.3,44.3,32.8,28.2,28.0,27.9. delta LCMS found m/z: 757.7(M + H),779.6(M-H + Na).

Example 4: synthesis of Compound 8

Compound 8 was prepared according to the previously reported procedure as described below (see, e.g., Kielar et al, J. Am. chem. Soc. 2010,132: 7836-7837).

Step 1: (R) -2- (4,7, 10-Tris (carboxymethyl) -1,4,7, 10-tetraazacyclododecan-1-yl) glutaric acid (Compound 7)

A solution of compound 6(250mg, 0.33mmol) in TFA/DCM (4mL/2mL) was stirred for 18 h. The solvent was removed under reduced pressure and the above procedure was repeated until deprotection was complete to give compound 7 (M/z (ES +): 477.4(M + H), monitored by LCMS) which was used in the next step without further purification.

Step 2: synthesis of Compound 8

Mixing GdCl ₃ ·6H ₂ O (135mg, 0.36mmol) was added to a solution of Compound 7 in water (10 mL). The pH of the solution was adjusted to 5.5-6.0 by adding 1M NaOH solution until the pH stabilized. The reaction was heated at 50 ℃ overnight. After cooling, the reaction was adjusted to pH-10.9 by addition of 1M HCl, stirred for 40 minutes, and then centrifuged. The supernatant was adjusted to pH 6.5 and lyophilized to give compound 8 as a white solid. LCMS found m/z (ES-): 630.2(M),652.1(M-H + Na).

Example 5: synthesis of mcMPO-Gd

To a solution of compound 8(125mg, 0.2mmol) and triethylamine (56 μ L, 0.4mmol) in DMSO (3mL) were added edc.hcl (57mg, 0.3mmol) and HOBt (40mg, 0.3 mmol). After 20 min, a solution of intermediate 4(76mg, 0.2mmol) in DMSO was added to the reaction. The reaction mixture was stirred at room temperature for 1 hour and then subjected to preparative HPLC to give the desired mcMPO-Gd as a white powder (25%). LCMS found m/z (ES +): 992.5(M + H); HRMS: 992.2795(M + H, calculated C) ₄₀ H ₄₉ GdN ₈ O ₁₂ ,992.2793)

Example 6: stability of mcMPO-Gd: with Zn ²⁺ Transfer metallization of

Due to Zn ²⁺ Is the main competing ion for gadolinium and is present in high concentrations in blood, with mcmPO-Gd in the presence of Zn ² ⁺ The kinetic stability in the case of (a) is compared with that of other agents (see, e.g., Rodriguez et al, J. Am. chem., 2010,132: 168-.

Applying mcMPO-Gd or MPO-Gd (1mM) with ZnCl ₂ (2.5mM) PBS was incubated at 40 ℃ and the relaxation rate (R1) was measured at 0.47T (20MHz) at 40 ℃ for 95 hours using an inversion recovery pulse sequence on Bruker Minispec (Bruker analysis, North Billerica, Mass.). By kinetic index (R1) _(t) /R1( _t＝0 ) 0.8) and thermodynamic index (R1) _{(72 hours)} /R1 _(t＝0) ) The stability of the mcMCP-Gd is evaluated and compared with MPO-Gd (see, e.g., Rodriguez et al, J. Am. chem., 2010,132:168-177) and BMA-Gd-DTPA (see, e.g., Moumne et al, J. Organische., 2006,71: 3332-3334).

Zn ²⁺ With Gd ³⁺ The transfer metallization between the two will be due to GdPO ₄ The formation of a precipitate leads to a relaxation change. The T1 relaxation times were measured at different time points as previously described (see, e.g., Rodriguez et al, J. Am. chem., 2010,132: 168-177). As shown in Table 1, mcmPO-Gd had no T1 change within 95 hours and was defined as R1 _(t) /R1 _(t＝0) Long term index of>0.97 (t-72-95 h), where the ratio index>5,000 minutes. This long-term index is significantly higher than the long-term index of MPO-Gd (0.76) and the long-term index of BMA-Gd-DTPA (0.09) (see, e.g., Laurent et al, radiology research, 2001,36: 115-.

Table 1.

	R1 _{(t 72 hours)} /R1 _(t＝0)	For R1 _(t) /R1 _(t＝0) T (min) ═ 0.8
			mcMPO-Gd	>0.97	>5000
MPO-Gd	0.76	2866
			BMA-DTPA-Gd	0.09	50-60

Example 7: relaxation potency of mcMPO-Gd

Relaxation efficiency (r) of MRI contrast agents ₁ ) Reflecting the sensitivity of the agent. The relaxivity of mcMPO-Gd containing PBS at concentrations of 0.33mM, 0.5mM, 0.75mM and 1mM was measured at 40 ℃. The 1/T1 values were plotted against the mcMPO-Gd concentration and fitted with linear regression. The slope value of the linear function is defined as the relaxation efficiency of the mcMPO-Gd (see e.g. Rohrer et al, radiology research 2005,40: 715-724). The relaxation potency of mcMPO-Gd at 0.47T (PBS, 40 ℃) was 5.4mM ^-1 s ^-1 As shown in fig. 2A, which is slightly higher than the previous phase4.3mM determined for MPO-Gd under the same conditions ^-1 s ^-1 (see, e.g., Querol et al, organic bulletin 2005,7: 1719-.

Example 8: in vitro Activity of mcMPO-Gd following activation by MPO

To compare the reactivity of mcMPO-Gd and MPO-Gd with MPO, a solution of mcMPO-Gd or MPO-Gd in PBS (0.5mM, total volume 150. mu.L) was mixed with glucose (as H) ₂ O ₂ A donor; 6 μ L, 1M), glucose oxidase (GOX; 4. mu.L, 1mg/mL) and MPO (10. mu.L, 2mg/mL) were incubated together at 40 ℃. The reaction was stopped by the addition of sodium azide (1 μ L, 250mg/ml) and the T1 relaxation times at 0 (before addition of MPO), 1 min, 2 min, 5 min, 10 min, 30 min, 60 min, 120 min and 180 min were measured at 40 ℃. The R1 ratio is expressed as (R1) _(t) -R1 _(t＝0) )/R1 _(t＝0) 。

As shown in FIG. 2B, the T1 change was more than 3-fold greater than the MPO-Gd change over 3 hours for mcMPO-Gd, indicating a higher activation efficiency for mcMPO-Gd than MPO-Gd.

Example 9: binding to proteins

The ability of mcMPO-Gd and MPO-Gd to bind to Bovine Serum Albumin (BSA) after activation by MPO was evaluated. A solution of mcMPO-Gd/MPO-Gd (0.5mM) in PBS (total volume 150. mu.L) was incubated with glucose (6. mu.L, 1M), GOX (4. mu.L, 1mg/mL) and MPO (10. mu.L, 2mg/mL) with/without BSA at 40 ℃ for 1 hour. The relaxation rates were measured as described above.

As shown in Table 2, MPO-Gd showed a modest improvement of 22% (32% to 54%) in T1 shortening in the presence of BSA. However, in the presence of BSA, mcMPO-Gd showed an improvement of 41% (80-121%). Overall, MPO-mediated mccmpo-Gd activation was 2.2-fold more potent in binding in the presence of BSA than MPO-Gd. These data indicate that mcMPO-Gd responds more strongly to MPO activation than MPO-Gd.

Table 2.

Example 10: cytotoxic MTT assay

Cytotoxicity of mcMPO-Gd was assessed using RAW 264.7 cells and measured using a 3- (4, 5-dimethylthiazol-2-yl) 2, 5-diphenyl-tetrazolium bromide (MTT) reduction assay as described previously (e.g., Rodriguez et al, journal of the american chemical society, 2010,132: 168-. About 1.5X 10 per well in a 96-well plate ⁴ The individual cells were cultured at 37 ℃ in a solution of mcMPO-Gd containing 10% FBS and 5% DMSO in DMEM at 0mM, 0.1mM, 0.5mM, 1mM, 2mM and 5mM for 16 hours. After incubation, cells were treated with 100 μ L of 0.5mg/mL MTT for 2 hours at 37 deg.C, then 100 μ L of DMSO was added to each well, the plates were incubated overnight at 37 deg.C, and the optical density was measured at 570nm using a microplate reader (Tecan Safire2, Tecan corporation of Dov, Switzerland). No significant cytotoxicity was observed within 5mM, as shown in figure 3, which is much higher than the expected in vivo concentration.

Example 11: magnetic Resonance (MR) imaging of Complete Freund's Adjuvant (CFA) inflammation

The efficacy of mcMPO-Gd was demonstrated in a well established mouse subcutaneous inflammation model induced by Complete Freund's Adjuvant (CFA). Six to ten weeks old female C57BL/6J mice (Jackson Laboratories, Bar Harbor, ME) and MPO deficient mice were used for this experiment. Under isoflurane anesthesia, one forelimb of the mouse was injected subcutaneously with 1:1(v: v) CFA: PBS emulsion (total volume of 40. mu.L). PBS (40. mu.L) was injected on the other side as a control. After 24 hours, gadolinium agents (mcMPO-Gd containing PBS containing 10% DMSO and 20% N, N-dimethylacetamide; or MPO-Gd containing PBS containing 5% DMSO; or Dotarem containing PBS) were administered by tail vein injection and mice were imaged using continuous T1 weighted imaging with chemical fat suppression and respiratory gating (TR: 900 ms, TE: 13.59 ms, 0.156 × 0.156 × 0.5mm voxels) on a 4.7T small animal MR scanner (Bruker, Cambridge, MA) with orthogonal volume coils of 3cm at 0 min, 15 min, 30 min, 45 min and 60 min (Rapid MR International, Germany)). Attention is paid toThe Region (ROI) was manually drawn by a person who did not know the identity of the mouse or imaging agent used, and the contrast to noise ratio (CNR) was calculated as CNR ═ (SI) _{Pathological changes} -SI _{Skeletal muscle} )/SD _Background 。

24 hours after injection of the CFA emulsion into the shoulder of the mouse, a contrast agent (0.3mol/kg) was injected through the tail vein, and the animal was imaged. FIGS. 4A-4D show the administration of mcMPO-Gd, Dotarem, and MPO-Gd contrast agents before imaging and in wild-type mice, respectively, and at MPO gene deficiency (Mpo) ^–/– ) Representative images obtained 60 minutes after administration of a contrast agent of mcMPO-Gd in mice. In wild type mice, the contrast to noise ratio (CNR) observed with mcMPO-Gd increased within 60 minutes, while the CNR observed with Dotarem decreased rapidly over time, as shown in fig. 4E, indicating that mcMPO-Gd but not Dotarem was retained in inflammatory tissues, which may be the result of activation of inflammation-associated MPO activity prior to subsequent binding to protein tyrosine residues. In contrast, Mpo was treated with mcMPO-Gd ^–/– MR imaging of mice showed little increase in signal, as shown in fig. 4C and 4E, indicating the specificity of the signal observed with mcMPO-Gd to MPO in this model. The activation ratio of mcmmpo-Gd (AR ═ CNR (post-radiography)/CNR (post-first radiography)) increased compared to the 2-fold increase seen with MPO-Gd>4-fold (see, e.g., Breckwoldt et al, Proc. Natl. Acad. Sci. USA, 2008,105: 18584-.

Example 12: MR imaging of Tandem Stenosis (TS) and interventions with AZM198

It has been reported that MPO activity is elevated in unstable plaques compared to stable plaques in a Tandem Stenosis (TS) mouse model, as assessed by the conversion of ethidium hydride (hydroethidine) to 2-chloroethylene by MPO-Gd MRI and LC-MS/MS assays (see, e.g., Rashid et al, J. European Heart, 2018,39: 3301-3310).

Male apolipoprotein E gene deficiency (Apoe) ^–/– ) Mice (6 weeks old) were fed a Western Diet (Western Diet, WD) containing 22% fat and 0.15% cholesterol(SF00-219, Specialty Feeds, Western Australia) for a total of 13 weeks. Six weeks after the onset of WD, Tandem Stenosis (TS) was introduced into mice as previously described (see, e.g., Rashid et al, European journal of the Heart, 2018,39: 3301-3310). Anesthesia of Male Apoe with 4% Isofluoroether ^–/– A mouse. The right common carotid artery was cut from the annular connective tissue. Two stenoses were placed by tying a 6-0 blue braided polyester suture (TICRON 0.7Metric) around the exposed artery, with the distal stenosis 1mm from the carotid bifurcation and the proximal stenosis 3mm from the distal stenosis. Blood flow was measured before and after addition of each ligature using a perivascular module (Transonic, TS420) and a0.7 mm perivascular probe (Transonic MA0.7PSB). The flow per ligature in the TS is defined as 70% of the baseline flow after addition of the distal ligature and 20% of the baseline flow after addition of the proximal ligature. Changes in flow predispose the right carotid artery to atherosclerotic plaque, with a segment proximal to the proximal suture having an unstable phenotype characterized by continued thinning of the fibrous cap, massive inflammatory cells, occasional neovascularization, cap rupture and intra-plaque hemorrhage, and luminal thrombosis with fibrin and platelet deposition (see, e.g., Chen et al, circulation research 2013,113: 252-. In contrast, atheroma in the head-arm trunk contains thick caps and abundant collagen, and is characterized by a stable plaque phenotype (see, e.g., Rashid et al, european journal of the heart, 2018,39: 3301-3310).

Isoflurane anesthetized mice were imaged in prone position before surgery and at 1,2,4 and 7 weeks post TS surgery using a 9.4T Bruker Biospec 94/20Avance III system (Bruker, Ettlingen, Germany) with 35mm orthogonal radio frequency coils and respiratory gated image acquisition before and after intravenous administration of 0.3mmol/kg MPO-Gd through the tail vein catheter as previously described. T1 weighted fast spin echo (TurboRARE, T1-TSE) was obtained with the following parameters: TR 1500 ms, TE 8.5 ms, ETL 8, slice thickness 1mm, FOV 20X 20mm, matrix size 192X 192, in-plane resolution 104X 104 μm. This T1-TSE protocol was then repeated in a series of scans covering a period of one hour after the contrast agent injectionTo assess contrast agent influx and retention. OsiriX (version 10.0.2, Pixmeo, Switzerland) was used for image analysis. In the T1-TSE image, different regions of interest are assigned to the vessel wall, skeletal muscle (reference) and background (air). The contrast to noise ratio (CNR) is calculated as follows: CNR ═ (SI) _{Vessel wall} -SI _{Skeletal muscle} )/SD _Background . In addition to 1-2 serial sections in the brachiocephalic trunk (stable plaque), the mean CNR of three serial sections of plaque with unstable phenotype and corresponding segment of the left carotid artery (no plaque) was calculated. By calculating Δ CNR ═ CNR _{After the production of images} -CNR _{Before radiography} To evaluate the segment enhancements attributable to MPO.

For the pharmacological inhibition of MPO, AZM198 (AstraZeneca, Sweden) was administered by incorporating WD at a daily dose of 500. mu. mol/kg body weight, based on the average daily food consumption per mouse of-3.7 g as described previously (see, e.g., Rashid et al, European journal of the Heart, 2018,39: 3301-3310). AZM198 treatment was initiated 1 week after TS surgery and continued for 6 weeks, before and after intravenous administration of 0.1mmol/kg of mcMPO-Gd according to the protocol described above, until MRI was performed 7 weeks after TS surgery.

Imaging was performed 7 weeks after TS surgery by administering 0.3mmol/kg MPO-Gd or 0.1mmol/kg mcMPO-Gd via the tail vein. As shown in fig. 5A, 5B and 5D, the signal enhancement of mccmpo-Gd and MPO-Gd in unstable plaques (arrows) was comparable (Δ CNR 17.8 versus 15.0, P ═ 0.368), but the dose of mccmpo-Gd was three times lower.

To confirm that MR imaging of mcMPO-Gd can reflect MPO activity, TS mice were treated with MPO inhibitor AZM198 1 week post TS surgery and subjected to MRI after an additional 6 week treatment period (total 7 weeks post TS). After administration of mcMPO-Gd, CNR of stable and unstable plaques increased at 30 min after contrast agent administration, but subsequently (60 min) decreased in mice treated with AZM198, while not in control animals, as shown in fig. 6. Thus, Δ CNR in the region corresponding to unstable plaques was reduced by about 50% in AZM 198-treated mice compared to that seen in control mice, as shown in fig. 5C and 5E, indicating that AZM198 effectively inhibits MPO activity.

The results described in the examples show that mcMPO-Gd has superior sensitivity and efficacy both in vitro and in vivo compared to MPO-Gd. Because the activated portions are similar, and without being bound by theory, this difference may be the result of the two activated portions being closer together and their rigidity. In MPO-Gd, each activating moiety is attached to one side of DTPA, which can lead to binding of both proteins or itself after oxidation of MPO. However, in mcMPO-Gd, the close proximity of the two active moieties prevents self-binding and may force the agent to bind to the same protein. This increases the efficiency of protein binding (as shown in table 2) and further reduces the segment motion of multiple proteins bound to the agent. It is believed that due to the stability of the mccmpo-Gd and the enhanced ability to image destructive inflammation, the mccmpo-Gd will be able to detect and map destructive inflammation at lower doses in a number of diseases, including but not limited to multiple sclerosis, non-alcoholic steatohepatitis (NASH), rheumatoid arthritis and atherosclerosis. These data indicate that mcMPO-Gd is a candidate molecular imaging agent for conversion to human use.

Example 13: molecular docking experiments

To explore why mcMPO-Gd is more effective in detecting MPO activity than MPO-Gd, molecular docking studies were performed using AutoDock Vina (see, e.g., Trott et al, journal of computational chemistry (j.comp.chem.) 2010,31(2),455-61) and visualized by PyMol. Since the parameters of gadolinium (Gd) are not included in AutoDock Vina, Gd was replaced with iron (Fe) in docking experiments (see, e.g., Wu et al, journal of molecular models (j.mol. model), 2016,22(7), 161). Gd and Fe have the same positive charge; without being bound by theory, it is hypothesized that replacing Gd with Fe should not affect the docking results, since Gd is "buried" within the chelating moiety of the agent, rather than interacting directly with dockerin.

Unlike MPO inhibitors that block the active site, the 5-hydroxyindole portion of mcMPO-Gd is an MPO substrate that undergoes a single electron transfer with the compound I form of MPO in the MPO catalytic cycle and does not remain at the active site (see, e.g., Rodriguez et al, journal of american chemical society, 2010,132(1), 168-77). MPO-cyanide (MPO-CN) complexes have been suggested as a useful alternative to the study of compounds I, which have a short lifetime, since both compounds I and MPO-CN complexes contain six-coordinated low spin s ═ 1 iron centers; thus, the eutectic structure of MPO-CN complexes from human MPO-cyanide-thiocyanate (MPO-CN-SCN, PDB database: 1DNW) was selected as a model for docking experiments (see, e.g., Malle et al, British journal of pharmaceutics (Br.J. Pharmacol.) 2007,152(6), 838-54; Blair-Johnson et al, Biochemistry (Biochemistry) 2001,40(46), 13990-7; Hallingback et al, Biochemistry 2006,45(9), 2940-50).

In this study, twenty docking conformations were generated for each agent. As shown in fig. 7, the optimal conformation of mccmpo-Gd binding to MPO reveals the formation of hydrogen bonds between the two amide bonds, which limits the rotation of mccmpo-Gd by forming a five-membered ring, thereby increasing the rigidity of the agent. As shown in FIG. 8, it was found that the binding affinity of the 20 patterns of generated mcMPO-Gd from the docking simulation to MPO was generally lower than that of MPO-Gd (11.7-9.5kcal/mol versus 9.9-7.9 kcal/mol). Without being bound by theory, these effects may explain the higher efficacy of mcMPO-Gd when activated by MPO.

Visualization of the binding of MPO-Gd and mcMPO-Gd to MPO indicates that for both agents, one of the 5-hydroxyindole moieties parallels MPO heme, forming pi-pi stacking interactions, consistent with previous docking studies on serotonin and MPO (see, e.g., Hallingback et al, biochemistry 2006,45(9), 2940-50). However, in mcMPO-Gd, the hydroxyl group pointing to the heme center forms a hydrogen bond with cyanides (violet and blue), and the imine group interacts electrostatically with the carbonyl group from heme propionate (NH... O ═ C,

). The other 5-hydroxyindole moiety is perpendicular to the heme, in which the imine is separately from Pro ¹⁰¹ And Thr ¹⁰⁰ Form two hydrogen bonds (see FIG. 9, left)

In contrast, as shown in FIG. 9 (right), for MPO-Gd, 5-hydroxyOne of the indole moieties interacts with MPO heme without electrostatic interaction, while the other 5-hydroxyindole moiety interacts only with Phe ¹⁴⁷ A hydrogen bond is formed. These differences lead to an improved binding affinity of mcMPO-Gd compared to MPO-Gd. Therefore, an improved mechanism of mcMPO-Gd over MPO-Gd was also identified by molecular docking experiments.

Detection and differentiation of harmful and beneficial inflammation is challenging, but they are useful in view of their different roles in disease pathogenesis. MPO has been found to be an important marker of destructive inflammation, and the examples provided herein describe gadolinium-based activatable macrocyclic MPO-specific agents mcMPO-Gd that are significantly more stable than linear MPO-Gd agents and respond up to three times higher to MPO activity than previous MPO-sensing MRI agents.

OTHER EMBODIMENTS

It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

1. A compound of formula I:

or a pharmaceutically acceptable salt thereof, wherein:

ring a is selected from the group consisting of: c _6-10 Aryl, 4-16 membered heterocycloalkyl and 5-16 membered heteroaryl;

R ¹ including chelating groups and metals;

m is 0,1, 2, 3 or 4; and is

n is 0,1, 2, 3 or 4.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein ring a is selected from the group consisting of: phenyl, bicyclic 8-to 16-membered heterocycloalkyl, tricyclic 8-to 16-membered heterocycloalkyl, bicyclic 8-to 16-membered heteroaryl, and tricyclic 8-to 16-membered heteroaryl.

3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein ring a is selected from the group consisting of:

and

wherein

Represents rings A and L ¹ A bond between them.

4. The compound of any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein m is 1 or 2.

5. The compound of any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, wherein m is 1.

6. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein each R ² Independently selected from the group consisting of: OH, OCH ₃ 、C(O)CH ₃ And OC (O) CH ₃ 。

7. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein each R ² Is OH.

8. The compound of any one of claims 1, 6 and 7, or a pharmaceutically acceptable salt thereof, wherein ring a is selected from the group consisting of:

and

wherein

Represents rings A and L ¹ A bond between them.

9. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein ring a is selected from the group consisting of:

and

wherein

Represents rings A and L ¹ A bond between them.

10. The compound of any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, wherein ring B is selected from the group consisting of: phenyl, bicyclic 8-16 membered heterocycloalkyl, tricyclic 8-16 membered heterocycloalkyl, bicyclic 8-16 membered heteroaryl and tricyclic 8-16 membered heteroaryl.

11. The compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, wherein ring B is selected from the group consisting of:

and

wherein

Represents rings B and L ¹ A bond between them.

12. The compound of any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, wherein n is 1 or 2.

13. The compound of any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, wherein n is 1.

14. The compound of any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, wherein each R ³ Independently selected from the group consisting of: OH, OCH ₃ 、C(O)CH ₃ And OC (O) CH ₃ 。

15. The compound of any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, wherein each R ³ Is OH.

16. The compound according to any one of claims 1 to 11, 14 and 15, or a pharmaceutically acceptable salt thereof, wherein ring B is selected from the group consisting of:

and

wherein

Represents rings B and L ¹ A bond between them.

17. The compound of any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, wherein ring B is selected from the group consisting of:

and

wherein

Represents rings B and L ¹ A bond between them.

18. The compound of any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, wherein ring a and ring B are the same.

19. The compound of any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, wherein ring a and ring B are different.

20. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein ring a and ring B are each

21. The compound of any one of claims 1 to 20, or a pharmaceutically acceptable salt thereof, wherein L ¹ Is L ² -NHC(O)-L ² -NHC(O)-L ² -, and each L ² Is independently selected C _1-4 An alkylene group.

22. The compound of any one of claims 1 to 20, or a pharmaceutically acceptable salt thereof, wherein L ¹ Comprises the following steps:

wherein:

represents L ¹ A bond to ring a;

represents L ¹ A bond to ring B; and is

-represents L ¹ And R ¹ A bond between them.

23. The compound of any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof, wherein R ¹ The metal of (a) is selected from the group consisting of: gd (Gd) ³⁺ 、Mn ²⁺ 、 ⁶⁸ Ga、 ⁶⁴ Cu and ¹¹¹ In。

24. the compound of any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof, wherein R ¹ Said metal of (A) is Gd ³⁺ 。

25. The compound of any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof, wherein R ¹ The chelating group of (a) is selected from the group consisting of:

and

wherein M is said metal and represents R ¹ And L ¹ A bond between them.

26. The compound of any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof, wherein R ¹ Comprises the following steps:

wherein-represents R ¹ And L ¹ A bond between them.

27. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound of formula I is a compound of formula II:

or a pharmaceutically acceptable salt thereof.

28. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound of formula I is a compound of formula III:

or a pharmaceutically acceptable salt thereof.

29. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound of formula I is a compound of formula IV:

or a pharmaceutically acceptable salt thereof.

30. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound of formula I is a compound of formula Vb:

or a pharmaceutically acceptable salt thereof, wherein M is the metal.

31. The compound of claim 1, which is:

or a pharmaceutically acceptable salt thereof.

32. A pharmaceutical composition comprising a compound according to any one of claims 1 to 31, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier.

33. A method of imaging a cell or tissue sample, the method comprising:

i) administering to a subject a compound according to any one of claims 1 to 31, or a pharmaceutically acceptable salt thereof;

iii) imaging the cell or tissue sample using an imaging technique.

34. A method of imaging a liver of a subject, the method comprising:

i) administering to the subject a compound of any one of claims 1 to 31, or a pharmaceutically acceptable salt thereof;

iii) imaging the cell or tissue sample using an imaging technique.

35. The method of claim 34, wherein the subject has been identified as having non-alcoholic steatohepatitis.

36. A method of diagnosing a disease or disorder associated with abnormal myeloperoxidase activity in a subject, the method comprising:

iii) imaging the cell or tissue using imaging techniques.

37. The method of claim 36, wherein the method further comprises imaging the subject prior to step i).

38. A method of imaging myeloperoxidase activity in a cell, the method comprising:

i) contacting the cell with a compound of any one of claims 1-31, or a pharmaceutically acceptable salt thereof; and

ii) imaging the cell with an imaging technique.

39. A method of detecting myeloperoxidase activity in a cell or tissue sample, the method comprising:

i) contacting the cell or tissue sample with a compound of any one of claims 1-31, or a pharmaceutically acceptable salt thereof; and

ii) imaging the cell or tissue sample with an imaging technique.

40. A method of detecting myeloperoxidase activity in a subject, the method comprising:

i) administering to the subject a compound of any one of claims 1 to 31, or a pharmaceutically acceptable salt thereof; and

ii) imaging the subject with an imaging technique.

41. A method of monitoring treatment of a disease or disorder associated with abnormal myeloperoxidase activity in a subject, the method comprising:

ii) imaging the subject with an imaging technique;

iv) imaging the cells or tissues of the subject with an imaging technique; and

v) comparing the image of step i) with the image of step iv).

42. The method of claim 41, wherein the method further comprises administering to the subject a compound according to any one of claims 1-31, or a pharmaceutically acceptable salt thereof, after the administering of step iii) and before the imaging of step iv).

43. The method of any one of claims 33 to 42, wherein the imaging technique is selected from the group consisting of magnetic resonance imaging and nuclear imaging.

44. The method of any one of claims 36, 37, and 41-43, wherein the disease or disorder associated with abnormal myeloperoxidase activity is selected from the group consisting of: non-alcoholic steatohepatitis, cancer, rheumatic diseases, infectious diseases, central nervous system diseases, cardiovascular disorders, autoimmune disorders, and inflammation associated with one or more of cancer, rheumatic diseases, infectious diseases, central nervous system diseases, cardiovascular disorders, and autoimmune disorders.

45. The method of claim 44, wherein the central nervous system disorder is selected from the group consisting of: alzheimer's disease, stroke, epilepsy, Parkinson's disease, neurodegenerative diseases, and inflammation associated with one or more of Alzheimer's disease, stroke, epilepsy, Parkinson's disease, and neurodegenerative diseases.

46. The method of claim 44, wherein the cardiovascular disorder is selected from the group consisting of: atherosclerosis, myocardial infarction, atrial fibrillation, vasculitis, and inflammation associated with one or more of atherosclerosis, myocardial infarction, atrial fibrillation, and vasculitis.

47. The method of claim 44, wherein the autoimmune disorder is selected from the group consisting of: multiple sclerosis, meningitis, encephalitis, and inflammation associated with one or more of multiple sclerosis, meningitis, and encephalitis.

48. The method of claim 44, wherein the cancer is selected from the group consisting of: bladder cancer, breast cancer, carcinoma, cervical cancer, colorectal cancer, endometrial cancer, glioma, head and neck cancer, liver cancer, lung cancer, melanoma, ovarian cancer, pancreatic cancer, prostate cancer, renal cancer, skin cancer, stomach cancer, testicular cancer, leukemia, and thyroid cancer.

49. The method of claim 44, wherein the cancer is a solid tumor.

50. The method of claim 44, wherein the rheumatic disease is selected from the group consisting of rheumatoid arthritis, osteoarthritis, and inflammatory arthritis.

51. The method of claim 50, wherein the inflammatory arthritis is selected from the group consisting of gout and calcium pyrophosphate deposition disease (CPPD).

52. The method of claim 44, wherein the infectious disease is selected from the group consisting of a fungal disease and a bacterial disease.

53. The method of claim 44, wherein the disease or disorder associated with aberrant myeloperoxidase activity is non-alcoholic steatohepatitis.